Hi! After struggling with inclusion bodies in E. coli, we would like to try purifying our 6xHIS-tagged proteins from Saccharomyces cerevisiae using the ADH promoter for gene expression. Are specialized strains necessary (such as protease deficient strains), and if so, which publicly-available strains are recommended?
We were going to use the Pierce Y-PER product for cell lysis (instead of glass beads), but would appreciate any other reliable and convenient protocols you may have.
Thanks!
Brian Ayre
University of North Texas